Introduction to Transmission Electron Microscope

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Introduction to Transmission Electron Microscope

TEM micrographs of a bio-sample

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Introduction to Transmission Electron Microscope

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Introduction to Transmission Electron Microscope

Structure of an aperture

Structure of a magnetic lens

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Introduction to Transmission Electron Microscope
Interaction of electron beam and a thin specimen in TEM

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Introduction to Transmission Electron Microscope
Types of TEM image: Bright field (BF); Dark Field (DF) and Diffraction Pattern

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Introduction to Transmission Electron Microscope

Mechanisms of Image Formation

In TEM, the specimen is transparent for electrons; absorption of

electrons plays a minor role in image formation. Deflection mechanism
(scattering and diffraction of electrons) is mainly responsible for image
formation.

Mass-thickness contrast (biology and polymer specimens)

Diffraction contrast (major image formation mechanisms)

Phase contrast (high resolution image of crystal lattice)

* Resolution Power: 0.2 nm at 1 MeV with 20 nm thick sample

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Introduction to Transmission Electron Microscope
Mass-thickness contrast
During passage of an electron wave through a specimen, scattering of electrons
occurs at the atoms in this specimen. → A certain portion of the incident electrons is
deflected (scattered) in direction different from the primary beam direction

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Introduction to Transmission Electron Microscope
Mass-thickness contrast

The brightness of image is determined by intensity of electron beam leaving the lower
specimen surface and pass through the objective aperture.
I = I0 exp(-S*ρ∗t)
where: I is the transmitted beam intensity; Io the incident beam intensity; ρ∗t the mass
thickness ( ρ is the density and t is the thickness of the sample) and S the effective mass
scattering cross-section, including size of objective aperture and incident beam energy,
i.e. acceleration voltage.

The contrast, K, can be defined as

K = log (Io/I)

The factors affecting electron scattering:

• atomic number of specimen atoms
• thickness of specimen
• density of specimen
• opening of objective aperture (size of aperture)
• energy of electrons (acceleration voltage) HKUST
Introduction to Transmission Electron Microscope
Using a objective aperture can enhance the mass-thickness contrast

(a) (b)

An animal cell photographed (a) with and (b) without an objective aperture in position

* A reduced objective aperture limits angle of scattering collection

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Introduction to Transmission Electron Microscope
A reduced beam voltage can enhance the mass-thickness contrast

TEM micrographs of a PE thin film

Objective Aperture: 120 µm

at 60 kV

Objective Aperture: 120 µm

at 100 kV

Objective Aperture: 20 µm
at 100 kV HKUST
Introduction to Transmission Electron Microscope
Diffraction contrast

The diffraction phenominon in crystalline materials as

The constructive interference occurs when the Bragg equation is

obeyed.
nλ = 2d sin θ

The contrast is generated when the electron beam does not pass
through aperture due to the Bragg diffraction of crystalline
specimen.

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Introduction to Transmission Electron Microscope
Diffraction contrast

When the Intensity Ir is removed

from the primary beam Io by the
Bragg reflection, the image of the
respective sample area appears
dark. HKUST
Introduction to Transmission Electron Microscope
Examples of diffraction contrast

The grains and precipitate image of Al alloy (a)

Titling the specimen changes the contrast of grains (b)

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Introduction to Transmission Electron Microscope
Examples of diffraction contrast

TEM micrograph with diffraction

contrast of a semi-crystalline polymer
thin film. The amorphous and crystalline
phases are clearly seen.
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Introduction to Transmission Electron Microscope
Phase contrast
Both mass-thickness and diffraction contrast are based on the amplitude contrast
mechanisms because they employ only the amplitudes of the scattered waves.

Phase contrast results from interference between waves of different phases.

• Most electron scattering involve a phase change.

• Two or more diffracted beams are allowed to pass through the objective aperture.
• Each pair of beams, which interferes, will in principal give rise to a set of fringes
in the images.

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Introduction to Transmission Electron Microscope
Examples of phase contrast

Without phase contrast With phase contrast

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Introduction to Transmission Electron Microscope
Examples of phase contrast

Use of two (or more) scattered beams:-

•Forwarded beam
•Diffracted beam or
•Diffracted beams
Interference of the beams shows the lattice fringe
image of the material.

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Introduction to Transmission Electron Microscope

Techniques for final thinning:

Electro-polishing (for metals)

A specimen is made the anode in an electrolytic cell. When current is
passed, the specimen is dissolved from the anode.

Ion milling (for metal and ceramics)

A beam of energetic ions is directed at a solid. Atom can be knocked out
from specimen.

Ultra-microtomy (for polymers and biological samples)

A firmly mounted specimen is moved past a fixed knife of glass and
diamond.

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Introduction to Transmission Electron Microscope
TEM Sample preparation techniques
-- ultra-microtomy

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Introduction to Transmission Electron Microscope
TEM Sample preparation techniques
-- making of replicas for TEM

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Introduction to Transmission Electron Microscope
TEM Sample preparation techniques
-- shadowing process

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Introduction to Transmission Electron Microscope
TEM Sample preparation techniques
-- shadowing with metals

TEM micrograph of PE crystals, shadowed with Pt/Carbon

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Introduction to Transmission Electron Microscope
TEM Sample preparation techniques
RuO4 stained samples
-- staining with heavy metal oxides

OsO4 stained samples HKUST

Introduction to Transmission Electron Microscope

A TEM micrograph of a semi-crystalline thin film showing crystal structure HKUST

Introduction to Transmission Electron Microscope
A craze in Polystyrene (PS)

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Introduction to Transmission Electron Microscope

Comparison of selected characteristics of optical and electron microscopes

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